CN103788051A - Method of isolating epigallocatechin gallate from tea polyphenols - Google Patents
Method of isolating epigallocatechin gallate from tea polyphenols Download PDFInfo
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- CN103788051A CN103788051A CN201410072241.7A CN201410072241A CN103788051A CN 103788051 A CN103788051 A CN 103788051A CN 201410072241 A CN201410072241 A CN 201410072241A CN 103788051 A CN103788051 A CN 103788051A
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07D—HETEROCYCLIC COMPOUNDS
- C07D311/00—Heterocyclic compounds containing six-membered rings having one oxygen atom as the only hetero atom, condensed with other rings
- C07D311/02—Heterocyclic compounds containing six-membered rings having one oxygen atom as the only hetero atom, condensed with other rings ortho- or peri-condensed with carbocyclic rings or ring systems
- C07D311/04—Benzo[b]pyrans, not hydrogenated in the carbocyclic ring
- C07D311/58—Benzo[b]pyrans, not hydrogenated in the carbocyclic ring other than with oxygen or sulphur atoms in position 2 or 4
- C07D311/60—Benzo[b]pyrans, not hydrogenated in the carbocyclic ring other than with oxygen or sulphur atoms in position 2 or 4 with aryl radicals attached in position 2
- C07D311/62—Benzo[b]pyrans, not hydrogenated in the carbocyclic ring other than with oxygen or sulphur atoms in position 2 or 4 with aryl radicals attached in position 2 with oxygen atoms directly attached in position 3, e.g. anthocyanidins
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Abstract
The invention relates to the technical field of tea polyphenols extraction, and discloses a method of isolating epigallocatechin gallate from tea polyphenols. The method comprises the following steps that step 1, an acetylation reaction is conducted for the tea polyphenols by using acetylation reagents in a condition that a hydroxyl group is catalyzed by catalysts, and acetylated tea polyphenols is produced; step 2, acetylated epigallocatechin gallate is acquired from the acetylated tea polyphenols produced in the step 1 through a process of silica gel column chromatography and a process of eluent elution separation; step 3, the epigallocatechin gallate is acquired from the acetylated epigallocatechin gallate acquired from the step 2 through reduction and hydrolyzation by active metals in acid solvents. The epigallocatechin gallate from tea polyphenols produced through the method has the advantages of being good in color, high in yield, simple in operation steps, free in pollution, and prone to being applied in the production on a large scale.
Description
Technical field
The present invention relates to tea-polyphenol extractive technique field, in particular, a kind of particularly method that separates NVP-XAA 723 from tea-polyphenol.
Background technology
Tea-polyphenol has another name called tea tannin, tea tannin, it is the polyphenols extracting from tealeaves, comprise catechin, anthocyanidin, flavanol compound, hydroxyl flavanol compound, anthocyanin and glucoside unit, flavonols and phenolic acids etc., also contain the alkaloids substances such as catechu oxide compound and polycondensation product and coffee.In tea-polyphenol, content is the highest, activity is the strongest for NVP-XAA 723 (EGCG); there is very strong anti-oxidant activity because thering is special poly-hydroxy chemistry structure; can Cell protection and DNA avoid infringement, aspect anticancer and cardiovascular disorder, taking on important role.Research shows, EGCG has anti-oxidant significantly, remove interior free yl, antitumor, anti-mutation, anti-ageing, anti-inflammatory, a series of biological activity such as antiviral, all there are good action, these effects to make EGCG there is boundless application prospect and marketable value to preventing and treating malignant tumour, hyperlipidemia, arteriosclerosis, cerebral thrombosis, diabetes, obesity etc.But because catechin main ingredient structure is more similar with physicochemical property, and the existence of phenolic hydroxyl group in EGCG structure is allowed to condition at and is easy to oxidizedly in preparation process, want effectively from tea-polyphenol, to separate and obtain EGCG monomer and there is certain technical difficulty.Chan Tak Hang(W02007041891) invent the method for extracting catechin by acetylize from tealeaves, ten thousand raising of indices etc. have also been developed from tea-polyphenol with acetylize conversion and separation method and have been prepared monomer NVP-XAA 723 (China Patent Publication No. is CN101182319A), wherein, the deacetylation protective reaction of the full acetic ester of EGCG is the acid hydrolytic reaction carrying out in the acid solution of high density, the acid of having reacted rear high density allows the aftertreatment of reaction product and the recovery of solvent become very difficult, and in default of protection, the phenolic hydroxyl group that reaction produces is easily oxidized and product colour is deepened by airborne oxygen.
Summary of the invention
The object of the present invention is to provide a kind of method that separates NVP-XAA 723 from tea-polyphenol, the NVP-XAA 723 that adopts the method to produce has the advantages such as color and luster is good, yield is high, operation steps is simple, pollution-free.
In order to solve problem set forth above, the technical solution used in the present invention is:
A method that separates NVP-XAA 723 from tea-polyphenol, comprises the following steps:
The first step, tea-polyphenol is carried out under catalyst to acetylization reaction at hydroxyl with acetylation reagent, make full acetylated tea-polyphenol;
Second step, by the full acetylated tea-polyphenol making in the first step through silica gel column chromatography, eluent wash-out separate, obtain full acetylated NVP-XAA 723;
The 3rd step, the full acetylated NVP-XAA 723 making in second step is obtained to NVP-XAA 723 through active metal reductive hydrolysis in acid solvent.
According to a preferred embodiment of the invention: the ripple metal alive adopting in described the 3rd step is zinc or magnesium.
According to a preferred embodiment of the invention: the acetylation reagent adopting in the described the first step is diacetyl oxide or Acetyl Chloride 98Min., and the catalyzer adopting be DMAP.
According to a preferred embodiment of the invention: the silica gel of the silica gel column chromatography in described second step is 200-300 order.
According to a preferred embodiment of the invention: the eluent adopting in described second step is ethyl acetate and 6# solvent extraction oil mixt, the volume ratio of ethyl acetate and 6# solvent extraction oil is that volume ratio is 1:0.5-2 and in described mixing volume.
According to a preferred embodiment of the invention: the eluent adopting in described second step is ethyl acetate and dichloromethane mixture, and the volume ratio of described ethyl acetate and methylene dichloride is 1:3-9.
According to a preferred embodiment of the invention: the acid solvent adopting in described the 3rd step is the solvent regulating through non-oxidizing acid.
According to a preferred embodiment of the invention: described non-oxidizing acid is hydrochloric acid, Hydrogen bromide or acetic acid.
According to a preferred embodiment of the invention: described solvent is the mixed solvent of methyl alcohol, ethanol, ethyl acetate or methylene dichloride and water.
Compared with prior art, beneficial effect of the present invention is:
1, the present invention has utilized the reduction characteristic of ripple metal alive, obtain phenolic hydroxyl group being adjusted to reductive hydrolysis acetic acid phenolic ester in weakly acidic conventional solvent, and because the protection of the ripple metallic reducing effect of living, can make the phenolic hydroxyl group that hydrolysis produces be difficult for the oxidation stain by airborne oxygen institute, guarantee the yield of product;
2 and reaction conditions gentleness, at reaction solution after completion of the reaction through simply filtering, concentrated and crystallization can obtain white NVP-XAA 723, and have that color and luster is good, yield is high, operation steps is simple, free of contamination advantage, be easy to application on large-scale production.
Embodiment
Below in conjunction with embodiment, the present invention is described in further detail, but embodiments of the present invention are not limited to this.
Embodiment mono-:
Take commercially available tea-polyphenol (polyphenol content 98%, EGCG50%) 50g, DMAP (DMAP) 2g, put in flask, add 500ml acetic acid ethyl dissolution and stir, slowly dripping diacetyl oxide 20ml, stirring reaction 6 hours, sampling and react completely with TLC monitoring after, under agitation condition, add 0.2mol/L dilute hydrochloric acid 200ml washing, wash altogether 3 times, and then wash with 200ml purified water, separate after organic phase concentrating under reduced pressure at 60 ℃, obtain thick liquid.After thick liquid is dissolved with methylene dichloride, through silica gel column chromatography, the eluent of employing is dichloromethane/ethyl acetate (volume ratio V/V=4/1), separates and obtains the full acetylated epigallocatechin gallate solid of white.
Full acetylated epigallocatechin gallate solid, with 500ml, 95% dissolve with ethanol, is added to acetic acid 20ml, stir; heating in water bath to 35 ℃; add magnesium grain 25g, stirring reaction 8 hours, after TLC monitoring reacts completely; filter; concentrating under reduced pressure boils off solvent, enriched material after appropriate purified water is dissolved to 10 ℃ at cooling crystallization 2 hours, filter; vacuum-drying obtains NVP-XAA 723 (EGCG) white powder 21.5g, and yield is 86%.
Embodiment bis-:
Other are identical with embodiment mono-for the present embodiment, difference is: the silica gel column chromatography eluent of employing is ethyl acetate/6# solvent extraction oil (volume ratio V/V=1/1.5), after vacuum-drying, obtain NVP-XAA 723 (EGCG) white powder 19.3g, yield is 77.2%.
Embodiment tri-:
Other are identical with embodiment mono-for the present embodiment; difference is: full acetylated epigallocatechin gallate solid dissolves with 200ml methylene dichloride; add 0.2mol/L dilute hydrochloric acid 300ml; stir; heating in water bath to 35 ℃; add magnesium grain 25g; stirring reaction 8 hours; after TLC monitoring reacts completely, filter, concentrating under reduced pressure boils off solvent; enriched material after appropriate purified water is dissolved to 10 ℃ at cooling crystallization 2 hours; filter, vacuum-drying obtains NVP-XAA 723 (EGCG) white powder 22.1g, and yield is 88.4%.
Embodiment tetra-:
Other are identical with embodiment mono-for the present embodiment, and difference is: change magnesium grain into zinc powder, vacuum-drying obtains NVP-XAA 723 (EGCG) white powder 20.3g, and yield is 81.2%.
Above-described embodiment is preferably embodiment of the present invention; but embodiments of the present invention are not restricted to the described embodiments; other any do not deviate from change, the modification done under spirit of the present invention and principle, substitutes, combination, simplify; all should be equivalent substitute mode, within being included in protection scope of the present invention.
Claims (9)
1. a method that separates NVP-XAA 723 from tea-polyphenol, is characterized in that, the method comprises the following steps:
The first step, tea-polyphenol is carried out under catalyst to acetylization reaction at hydroxyl with acetylation reagent, make full acetylated tea-polyphenol;
Second step, by the full acetylated tea-polyphenol making in the first step through silica gel column chromatography, eluent wash-out separate, obtain full acetylated NVP-XAA 723;
The 3rd step, the full acetylated NVP-XAA 723 making in second step is obtained to NVP-XAA 723 through active metal reductive hydrolysis in acid solvent.
2. the method that separates NVP-XAA 723 from tea-polyphenol according to claim 1, is characterized in that: the ripple metal alive adopting in described the 3rd step is zinc or magnesium.
3. the method that separates NVP-XAA 723 from tea-polyphenol according to claim 1 and 2; it is characterized in that: the acetylation reagent adopting in the described the first step is diacetyl oxide or Acetyl Chloride 98Min., and the catalyzer adopting is DMAP.
4. the method that separates NVP-XAA 723 from tea-polyphenol according to claim 1 and 2, is characterized in that: the silica gel of the silica gel column chromatography in described second step is 200-300 order.
5. the method that separates NVP-XAA 723 from tea-polyphenol according to claim 1 and 2, it is characterized in that: the eluent adopting in described second step is ethyl acetate and 6# solvent extraction oil mixt, and in described mixing volume, the volume ratio of ethyl acetate and 6# solvent extraction oil is that volume ratio is 1:0.5-2.
6. the method that separates NVP-XAA 723 from tea-polyphenol according to claim 1 and 2, it is characterized in that: the eluent adopting in described second step is ethyl acetate and dichloromethane mixture, and the volume ratio of described ethyl acetate and methylene dichloride is 1:3-9.
7. the method that separates NVP-XAA 723 from tea-polyphenol according to claim 1 and 2, is characterized in that: the acid solvent adopting in described the 3rd step is the solvent regulating through non-oxidizing acid.
8. the method that separates NVP-XAA 723 from tea-polyphenol according to claim 7, is characterized in that: described non-oxidizing acid is hydrochloric acid, Hydrogen bromide or acetic acid.
9. the method that separates NVP-XAA 723 from tea-polyphenol according to claim 8, is characterized in that: described solvent is the mixed solvent of methyl alcohol, ethanol, ethyl acetate or methylene dichloride and water.
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| CN201410072241.7A CN103788051A (en) | 2014-02-28 | 2014-02-28 | Method of isolating epigallocatechin gallate from tea polyphenols |
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107057437A (en) * | 2017-04-06 | 2017-08-18 | 天长市润达金属防锈助剂有限公司 | A kind of antibacterial modified antirust agent of high resistant |
| CN107235948A (en) * | 2017-05-31 | 2017-10-10 | 湖北中鑫生物科技有限公司 | The guard method of phenolic hydroxyl group during a kind of Tea Polyphenols molecular modification |
| CN111250068A (en) * | 2020-01-21 | 2020-06-09 | 中华全国供销合作总社杭州茶叶研究所 | Application and method of acetylation medium filler for separating and purifying tea catechin |
-
2014
- 2014-02-28 CN CN201410072241.7A patent/CN103788051A/en active Pending
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107057437A (en) * | 2017-04-06 | 2017-08-18 | 天长市润达金属防锈助剂有限公司 | A kind of antibacterial modified antirust agent of high resistant |
| CN107235948A (en) * | 2017-05-31 | 2017-10-10 | 湖北中鑫生物科技有限公司 | The guard method of phenolic hydroxyl group during a kind of Tea Polyphenols molecular modification |
| CN111250068A (en) * | 2020-01-21 | 2020-06-09 | 中华全国供销合作总社杭州茶叶研究所 | Application and method of acetylation medium filler for separating and purifying tea catechin |
| CN111250068B (en) * | 2020-01-21 | 2022-10-18 | 中华全国供销合作总社杭州茶叶研究所 | Application and method of acetylation medium filler for separating and purifying tea catechin |
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Application publication date: 20140514 |